nha-oc/NHA2 overexpression prevents caspase activation in RAW 264.7 cells.
RAW 264.7 cells were transfected with pNHA-oc/V5-6X His. 48 hours later cells were stimulated with RANKL overnight (RANKL +, pNHA-oc +). Control cultures were unstimulated (RANKL -, pNHA-oc +). In parallel, cell were mock transfected and either RANKL-stimulated (RANKL +, pNHA-oc -) or not (RANKL -, pNHA-oc -). Three days after the transfection, protein and RNA was prepared and analyzed.
(A) RT-PCR analysis of transfected and mock transfected cells show nha-oc/NHA2 expression only in transfected cells.
(B) Protein extracts were subjected to Western Blot analysis using caspase-specific antibodies. Caspase activation is inhibited in cells expressing nha-oc/NHA2, regardless RANKL stimulation. An asterisk next to the molecular weight indicates that those bands represent the cleaved (not full length) caspase. The antibodies can detect both the full length and the cleaved products (with the exception of those against cleaved caspase 7 and PARP, that only detect the cleaved products)